Developing a single-objective lens, two-photon excitation, light-sheet microscopy (2P-SCAPE) for high-speed, volumetric imaging of biological tissues

Hang Yu

Two-photon microscopy has become a widely adopted tool for functional Calcium imaging in neuroscience research. Due to the decreased scattering at near-infrared wavelengths, two-photon excitation improves penetration depth and image contrast in the mouse brain over single-photon excitation. However, the imaging acquisition is usually performed in a laser-scanning approach, which restricts the system’s spatiotemporal bandwidth, allowing only a limited number of neurons to be captured from a 2D image plane. This dissertation focuses on the development of a single-objective lens, light-sheet excitation, two-photon microscopy approach (2P-SCAPE) that dramatically improves the system’s bandwidth over laser-scanning. The spatial multiplexing provided by light-sheet excitation resolved the trade-off between imaging speed and signal-to-noise ratio in laser-scanning. The single objective lens...